Optimization of Bacterial Whole Cell Bioreporters for Toxicity Assay of Environmental Samples
Optimization of Bacterial Whole Cell Bioreporters for Toxicity Assay of Environmental Samples
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DOI：
10.1021/es901349r
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发表时间：
2009-10-15
影响因子：
11.4
通讯作者：
Huang, Wei E.
中科院分区：
文献类型：
--
作者：
Song, Yizhi;Li, Guanghe;Huang, Wei E.
为了优化细菌全细胞生物传感器用于环境污染物检测，我们构建了一株毒性敏感菌株不动杆菌湾/YI ADP1_recA_lux。ADP1_recA_lux是一种基于染色体的生物报告器，它使传感系统稳定，并且不需要抗生素来维持特征。当ADP1_recA_lux暴露在DNA损伤毒物中时，它被激活以表达生物发光。由于ADP1recAlux恒定地表达生物发光的基线水平，因此避免了假阴性结果。宿主菌株A.bay/yi ADP1是自然环境中典型的水和土壤细菌的理想模式菌株，在活性、维持和储存方面比大肠杆菌更健壮。在广泛的温度范围(10-40℃)中，ADP1_recA_lux被用于检测各种有毒或潜在的有毒化合物，包括丝裂霉素C(MMC)、甲磺酸甲酯、溴化乙锭、过氧化氢、甲苯、单壁纳米碳管(SWNCT)、纳米金胶体(20 Nm)、芘、苯并[a]芘和紫外光。这些暴露表明ADP1recAlux能够定性和定量地检测到遗传毒性和细胞毒性。ADP1recAlux生物报告的最佳诱导时间为3h，对MMC和苯并[a]芘的检出限分别为1.5 nM和0.4 nM。ADP1recAlux还用于检测受多种酚类化合物污染的地下水的毒性，生物报告毒性检测结果与化学分析结果吻合较好。优化后的全细胞生物报告软件ADP1_recA_lux可为环境样品中的毒性检测提供一种简单、快速、稳定、定量、可靠、成本低廉的有效方法。
In a study to optimize bacterial whole cell biosensors (bioreporters) for the detection of environmental contaminants, we constructed a toxicity sensing strain Acinetobacter bay/yi ADP1_recA_lux. The ADP1_recA_lux is a chromosomally based bioreporter which makes the sensing system stable and negates the need for antibiotics to maintain the trait. The ADP1_recA_lux is activated to express bioluminescence when it is exposed to DNA damaging toxicants. Since the ADP1_recA_lux constantly expresses a baseline level of bioluminescence, false negative results are avoided. The host strain, A. bay/yi ADP1, is an ideal model strain typical of water and soil bacteria occurring in the natural environment and it is more robust than E. coli in terms of viability, maintenance, and storage. The expression of reporter genes - luxCDABE cloned from Photorhabdus luminescens - is robust in a broad range of temperature (10-40 degrees C), The ADP1_recA_lux was used to detect a variety of toxic or potentially toxic compounds including mitomycin C (MMC), methyl methanesulfonate, ethidium bromide, H2O2, toluene, single-wall nanocarbon tubes (SWNCT), nano Au colloids (20 nm), pyrene, beno[a]pyrene, and UV light. These exposures revealed that the ADP1_recA_lux was able to detect both genotoxicity and cytoxicity, qualitatively and quantitatively. The optimal induction time of the ADP1_recA_lux bioreporter was 3 h, and the detection limits for MMC and benezo[a]pyrene were 1.5 nM and 0.4 nM, respectively. The ADP1_recA_lux was also used to detect toxicity of groundwater contaminated by a mixture of phenolic compounds, and the bioreporter toxicity detection was in a good agreement with chemical analysis. The optimized whole cell bioreporter ADP1_recA_lux could be valuable in providing a simple, rapid, stable, quantitative, robust, and costly efficient approach for the detection of toxicity in environmental samples.